RT Journal Article
T1 Chemical Modification of a Dehydratase Enzyme Involved in Bacterial Virulence by an Ammonium Derivative: Evidence of its Active Site Covalent Adduct
A1 González Bello, Concepción
A1 Tizón, Lorena
A1 Lence Quintana, Emilio José
A1 Otero Casas, José Manuel
A1 Raaij, Mark J. van
A1 Martínez Guitián, Marta
A1 Beceiro Casas, Alejandro
A1 Thompson, Paul
A1 Hawkins, Alastair R.
AB The first example of an ammonium derivative that causes a specific modification of the active site of type I dehydroquinase (DHQ1), a dehydratase enzyme that is a promising target for antivirulence drug discovery, is described. The resolution at 1.35 Å of the crystal structure of DHQ1 from Salmonella typhi chemically modified by this ammonium derivative revealed that the ligand is covalently attached to the essential Lys170 through the formation of an amine. The detection by mass spectroscopy of the reaction intermediates, in conjunction with the results of molecular dynamics simulations, allowed us to explain the inhibition mechanism and the experimentally observed differences between S. typhi and Staphylococcus aureus enzymes. The results presented here reveal that the replacement of Phe225 in St-DHQ1 by Tyr214 in Sa-DHQ1 and its hydrogen bonding interaction with the conserved water molecule observed in several crystal structures protects the amino adduct against further dehydration/aromatization reactions. In contrast, for the St-DHQ1 enzyme, the carboxylate group of Asp114, with the assistance of this water molecule, would trigger the formation of a Schiff base that can undergo further dehydration reactions until full aromatization of the cyclohexane ring is achieved. Moreover, in vitro antivirulence studies showed that the reported compound is able to reduce the ability of Salmonella Enteritidis to kill A459 respiratory cells. These studies have identified a good scaffold for the design of irreversible inhibitors that can be used as drugs and has opened up new opportunities for the development of novel antivirulence agents by targeting the DHQ1 enzyme
PB American Chemical Society
SN 0002-7863
YR 2015
FD 2015
LK http://hdl.handle.net/10347/16933
UL http://hdl.handle.net/10347/16933
LA eng
NO González-Bello, C., Tizón, L., Lence, E., Otero, J., van Raaij, M., & Martinez-Guitian, M. et al. (2015). Chemical Modification of a Dehydratase Enzyme Involved in Bacterial Virulence by an Ammonium Derivative: Evidence of its Active Site Covalent Adduct. Journal Of The American Chemical Society, 137(29), 9333-9343. doi: 10.1021/jacs.5b04080
NO Financial support from the Spanish Ministry of Science andInnovation (SAF2013-42899-R), Xunta de Galicia (GRC2013-041), and the European Regional Development Fund (ERDF)is gratefully acknowledged. E.L. thanks the Xunta de Galicia forhis postdoctoral fellowship. A.B. thanks the Miguel ServetProgramme ISCIII-FEDER (CP13/00226) and the ISCIIIGeneralSubdirection of Assesment and Promotion of theResearch (PI14/00059) for financial support
DS Minerva
RD 28 abr 2026